Highly sensitive protein detection via covalently linked aptamer to MoS₂ and exonuclease-assisted amplification strategy

Figures & data

Figure 1 TEM image of AuNPs dispersed on the surface of MoS₂.

Abbreviations: AuNP, gold nanoparticle; MoS₂, molybdenum disulfide; TEM, transmission electron microscopy.

Figure 2 The fluorescence intensity of FAM-modified aptamer (10 nM) in the presence of various concentrations (0, 10, 15, 20, 25, 30, 40, and 50 μg/mL) of AuNPs@MoS₂ nanocomposites.

Abbreviations: AuNPs, gold nanoparticles; MoS₂, molybdenum disulfide; FAM, carboxyfluorescein.

Figure 3 (A) The fluorescence intensity of FAM-modified aptamer (10 nM) in the presence of AuNPs@MoS₂ nanocomposites (40 μg/mL) with varying concentrations of thrombin (0.03 pM, 0.04 pM, 0.05 pM, 0.06 pM, 0.4 pM, 0.6 pM, 0.8 pM, 1.2 pM, 4 pM, 5 pM, 60 pM, and 120 pM) and (0.03 UμL-1) Exo. (B) The values of [F/F₀-1] for assay with the concentration of thrombin are shown.

Note: All data were collected from three measurements, and the error bars indicate the standard deviation.

Abbreviations: AuNPs, gold nanoparticles; MoS₂, molybdenum disulfide; Exo, exonuclease; FAM, carboxyfluorescein; F₀, values of fluorescence intensities without thrombin; F, values of fluorescence intensities with thrombin.

Table 1 Comparison of different methods for thrombin detection

Detection technique	Linear range	Detection limit	References
Fluorescence	0.0048 nM–20 nM	4.8 pM	^Citation18
SPR	0.1 nM–75 nM	0.1 nM	^Citation22
Luminescence energy transfer	0.375 nM–11.25 nM	0.118 nM	^Citation24
Electrochemical sandwich assay	100 pM–100 nM	1.5 pM	^Citation26
Electrochemistry	5 pM–50 nM	0.9 pM	^Citation16
Strip biosensors	6.4 pM–500 nM	4.9 pM	^Citation27
UV-vis	1.3 nM–133 nM	0.61 nM	^Citation28
Electrochemistry	0.3 nM–50 nM	0.01 nM	^Citation25
Fluorescence	0.1 nM–10 nM	0.1 nM	^Citation29
SERS	0.27 pM–2.7 pM	0.27 pM	^Citation30
Fluorescence	0.04 pM–140 pM	6 fM	Present work

Abbreviations: SERS, surface-enhanced raman scattering; SPR, surface plasmon resonance; UV, ultraviolet; vis, visible.

Figure 4 The fluorescence intensity of FAM-modified aptamer (10 nM)-MoS₂ (40 μg/mL) in the presence of other proteins (BSA, IgG, and lysozyme) at the concentration of 0.001 nM.

Abbreviations: BSA, bovine serum albumin; Exo, exonuclease; FAM, carboxyfluorescein; IgG, immunoglobulin G; MoS₂, molybdenum disulfide.

Scheme 1 The strategy for the aptamer–MoS₂ biosensor based on FRET to detect thrombin.

Abbreviations: FRET, fluorescence resonance energy transfer; MoS₂, molybdenum disulfide; AuNPs, gold nanoparticles.

Table 2 Results for the determination of thrombin in human blood serum

Serum sample	Concentration of thrombin added (pM)	Concentration obtained with aptasensor (nM)	Recovery (%)	RSD (%)
1	1.2	1.002	83.51	7.01
2	10	9.209	92.09	5.32
3	100	103.02	103.02	3.76

Abbreviation: RSD, relative standard deviation.

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